天津医药

• 论著 •    

生物素亲和素系统在ECP的实验效能

赵俊芳1,王学谦2,李桂珍1   

  1. 1. 天津市中医药研究院附属医院
    2. 天津市天津医院
  • 收稿日期:2011-04-08 修回日期:2011-08-11 出版日期:2012-03-15 发布日期:2012-03-15
  • 通讯作者: 赵俊芳

The Application of ABS in Double-antibody Sandwich ELISA Method for Eosinophilia Cationic Protein

  • Received:2011-04-08 Revised:2011-08-11 Published:2012-03-15 Online:2012-03-15
  • Contact: Jun-Fang ZHAO

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摘要： 目的 评估生物素亲和素系统对ECP双抗夹心ELISA试验效能的影响。 方法 1. 生物素化抗-ECP单克隆抗体；2.双抗夹心ELISA ECP检测试剂盒的制备；3.制备先包被生物素的ELISA试剂盒；4.制备先包被亲和素的ELISA试剂盒；5.比较三种检测方法。 结果 1. 三种方法的比较：双抗夹心ELISA ECP检测试剂盒线性范围为2.5～200ng/ml，灵敏度为2.75ng/ml，相对偏差为11.5％；生物素亲和素包被法线性范围为2.5～500ng/ml，灵敏度为2.61ng/ml，相对偏差为10.9％；亲和素包被法线性范围为2.5～500ng/ml，灵敏度为2.68ng/ml，相对偏差为11.9％。2.三种方法的重复性没有显著性差异，稳定性亲和素包被法最为稳定，抗体包被法次之，生物素包被法最不稳定，抗体直接包被法操作最为简易，但抗体用量最多。 结论 生物素亲和素包被可提高实验的灵敏度、线性范围，减少抗体用量，节约抗体包被时间。

关键词: 嗜酸性粒细胞阳离子蛋白, 酶联免疫吸附试验, 生物素, 亲和素

Abstract: Objective Evaculate the effect of ABS on the test performance of the sandwich enzyme immunoassay of double antibodies kit (ELISA) for ECP . Methods 1. Biotinylation of monoclonal antibody against ECP. 2. The development of double antibodies ELISA kit for ECP. 3. The development of ELISA kit with biotin firstly coated. 4. The development of ELISA kit with avidin firstly coated. 5.The comparison of three methods. Result 1. The comparison of three methods:(1) The linear range、sensitivity and relative deviation of direct coating was 2.5~200 ng/ml, 2.75ng/ml ,11.5％，respectively. (2)Three factors of biotin-avidin coating test were 2.5~500 ng/ml、2.61ng/ml and 10.9％ ,respectively.(3) Three factors of avidin coating test was 2.5~500 ng/ml、2.68ng/ml and 11.9％ , respectively. 2.There was no significant difference among their reproducibility. Avidin coating test was the stablest method and antibody coating test was the simplest method, but its antibody dosage was the highest. Conclusion Biotin-avidin coating can elevate the linear range and sensitivity of the test and it could save antibody dosage and antibody coating time.

Key words: Key wordeosinophilia cationic protein, enzyme linked immunosorbent assay, biotin, avidin